sectioning

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Jeffrey,
I have always found that embryos in general do not section very nicely in
paraffin. I would recommend frozen sections or plastic sections for
anything less than 72 hours. The anatomy is preserved much better. If
your staining is robust, in situs following by sectioning is easier. If
not, you can do in situs on frozen sections but this is a bit more
tricky. I can send you a protocol if you decide this is necessary.
Also, it seems embedding quality varies widely due both to the fix and
also to whatever processor (or by hand) you might be using.

Mary, I would highly recommend the TRIZOL reagent from Gibco, followed
by cleanup with mini RNA columns from Qiagen. You can actually store
tissues in TRIZOL at -80. It is nice to have a mini-homogenizer, which
can be placed directly in the thawing TRIZOL. This procedure gives very
high quality RNA suitable for microarrays.

Melissa Haendel
ZFIN curation staff

[Only registered users see links. ] wrote:

happy to share my embedding protocol with you, please let me know. I have
not had vast experience yet with whole mount in situs, my experience
yielded me chewed up over digested skeletons, and not much else. I will
be revisiting this procedure in the near future, and any pearls of advice
would be greatly appreciated.
isolate adult zebra fish tissues for RNA isolation, is there anyone who
is doing this that I might be able to chat with?
is always an evil, never a good. We will not learn how to live together
in peace by killing each other's children. President Jimmy Carter
times there are problems with some of them. My most common problem is
that the embryos will often crumble and crack. I have tried isolating
the problem, but I can't seem to figure it out. Does anyone use
better for whole-mount in situ zebrafish? Or am I better off performing
in situ hybridization after sectioning? Any advice is appreciated.

--
Melissa Haendel, Ph.D.

ZFIN Scientific Curator

Zebrafish Information Network

5291 University of Oregon

Eugene, OR 97403-5291

Phone: (541) 346-5108


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